• Title/Summary/Keyword: In vitro activation

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Study on the Developmental Rate of In Vitro Cultured Cats Oocytes Recovered from Ovaries Collected at Different Stages of the Reproductive Cycle (번식주기의 단계별로 회수한 고양이 난자의 체외수정과 체외발생에 관한 연구)

  • 박상훈;이명헌;김무강;김상근
    • Journal of Embryo Transfer
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    • v.18 no.2
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    • pp.157-161
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    • 2003
  • The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes in vitro maturation of cats oocytes and development of IVM/IVF embryos. The results were summarized as follows : 1. The fertilization and developmental rate of fresh and salts-stored oocytes with and whithout cumulus cells were 65.7%, 17.1% and 28.6%, 8.6% and 57.1%, 13.3%, 23.3%, 3.3%, respectively. The rate of oocytes with cumulus cells(13.3%∼65.7%) was higher than that of denuded oocytes(3.3%∼28.6%). 2. The fertilization and developmental rate of oocytes recovered from ovaries collected at different stages of the reproductive cycle were 68.9%, 44.4%, 48.9% and 17.8%, 8.9%, 12.8%, respectively. 3. The fertilization and developmental rate of oocytes in vitro cultured at different time of incubation(24, 36 and 48 h) were 66.7%, 46.7%, 48.9% and 17.8%, 11.1%, 8.5%, respectively. respectively. The rate of oocytes incubated 24 h(66.7%) was higher than that oocytes incubated 36 and 48 h(46.7%∼48.9%). 4. The fertilization and developmental rate of oocytes treated activation and non-activation oocytes were 57.4%, 31.4% and 22.9%, 11.4%, respectively. The rate of oocytes treated activation was higher than that oocyte treat non-activation.

Surface Characteristics and Biocompatibility of Hydroxyapatite Deposited Ti alloys by Electrochemical Deposition

  • Lee, Kang;Choe, Han Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2015.11a
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    • pp.141-141
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    • 2015
  • In this study, a series of hydroxyaptite (HAp) are produced on Ti dental implant using electrochemical deposition. Based on the preliminary analysis of the coating structure, composition and morphology. In vitro studies were performed with MC3T3-E1 cell to investigate the effect of biological change on different surface conditions.

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Role of Poly (ADP-ribose) Polymerase Activation in Chemical Hypoxia-Induced Cell Injury in Renal Epithelial Cells

  • Jung Soon-Hee
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.441-446
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    • 2005
  • The molecular mechanism of ischemia/reperfusion injury remains unclear. Reactive oxygen species (ROS) are implicated in cell death caused by ischemia/reperfusion in vivo or hypoxia in vitro. Poly (ADP-ribose) polymerase (PARP) activation has been reported to be involved in hydrogen peroxide-induced cell death in renal epithelial cells. This study was therefore undertaken to evaluate the role of P ARP activation in chemical hypoxia in opossum kidney (OK) cells. Chemical hypoxia was induced by incubating cells with antimycin A, an inhibitor of mitochondrial electron transport. Exposure of OK cells to chemical hypoxia resulted in a time-dependent cell death. In OK cells subjected to chemical hypoxia, the generation of ROS was increased, and this increase was prevented by the $H_2O_2$ scavenger catalase. Chemical hypoxia increased P ARP activity and chemical hypoxia-induced cell death was prevented by the inhibitor of PARP activation 3-aminobenzamide. Catalase prevented OK cell death induced by chemical hypoxia. $H_2O_2$ caused PARP activation and $H_2O_2-induced$ cell death was prevented by 3-aminobenzamide. Taken together, these results indicate that chemical hypoxia-induced cell injury is mediated by PARP activation through H202 generation in renal epithelial cells.

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Carex pumila Extract Supresses Mast Cell Activation and IgE-Mediated Allergic Response in Mice (좀보리사초의 IgE 매개성 알레르기 반응 억제 효과 및 기전)

  • Lim, Hannah;Kim, Young Mi
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.356-362
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    • 2014
  • Allergic diseases have increased rapidly over the past decades, affecting an estimated 20~30% of the population in developed countries. In this study, we investigated whether or not a typical costal sand dune plant Carex pumila (CPE) suppresses the activation of mast cells and IgE-mediated allergic response in vitro and in vivo. As the results, the extract of Carex pumila inhibited antigen-stimulated degranulation in RBL-2H3 cells and Bone marrow-derived mast cells (BMMCs), and IgE-mediated passive cutaneous anaphylaxis (PCA) in mice. CPE also suppressed the production of pro-inflammatory cytokines, TNF-${\alpha}$ and IL-4, in antigen-stimulated mast cells. As its mechanism of action, CPE inhibited the activation of Syk in $Fc{\varepsilon}RI$-mediated signalling pathway, and that of LAT, a downstream adaptor molecule of Syk, in a dose-dependent manner. CPE also suppressed the activation of mitogen-activated protein (MAP) kinases, p38, ERK1/2, JNK, and Akt. Altogether, CPE inhibited mast cell activation and IgE-mediated allergic response by antigen through suppressing the activation of Syk. These results suggest that CPE may be useful for the treatment of allergic diseases.

Evaluation of Genotoxicity of CP Pharmacopuncture Using an In Vitro Chromosome Aberration Test in Chinese Hamster Lung Cell (Chinese Hamster Lung 세포를 이용한 염색체이상 시험을 이용한 CP약침의 유전독성평가)

  • Hwang, Ji Hye;Jung, Chul;Ku, Jaseung
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.34 no.6
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    • pp.355-361
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    • 2020
  • This study was designed to assess the toxicity of capsaicin-containing (CP) pharmacopunture using an in vitro chromosomal aberrations in Chinese hamster lung (CHL/IU) cells. In order to determine the high dose level in the main study of this study, a dose range finding study was conducted first. The high dose was selected at 10.0% of CP pharmacopuncture extract, and then diluted sequentially to produce lower dose levels of 5.00, 2.50, 1.25, 0.625 and 0.313% by applying a geometric ratio of 2. As a result, the cytotoxicity and precipitation of the CP pharmacopuncture as a test substance were not evident at any dose level during short-time treatment with and without metabolic activation and continuous treatment without metabolic activation. Therefore, the dose levels for this study were chosen as 10.0, 5.0, and 2.5%., and the treatment volume was 1.3 mL. In addition, negative and positive controls were set. In main study, the frequency of cells with chromosome aberrations in CP treated groups was less than 5% in short-time treatment with and without metabolic activation and continuous treatment without metabolic activation. In addition, there was no statistically significant difference when compared to the negative control group. The frequency of cells with structural chromosomal aberrations in the positive control group was more than 10% compared to the negative control group, and it increased statistically significantly. In conclusion, under the conditions of this study, CP pharmacopuncture did not show the possibility of causing chromosome aberrations.

Evaluation of the genetic toxicity of synthetic chemicals (V) -in vitro Chromosomal Aberration Assay with 17 chemicals in Chinese Hamster Lung Cells-

  • Ryu, Jae-Chun;Kim, Kyung-Ran;Kim, Youn-Jung;Choi, Hae-Yeon
    • Environmental Mutagens and Carcinogens
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    • v.22 no.4
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    • pp.215-222
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    • 2002
  • The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, to regulate and to evaluate the chemical hazard will be important to environment and human health. The clastogenicity of 17 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. Two most cytotoxic chemicals, dodecyl methacrylate (CAS No. 142-90-5) and 2-ethylhexyl methacrylate (CAS No. 688-84-6), among 17 chemicals tested revealed no clastogenicity in the range of 0.0165-0.066 $\mu\textrm{g}$/$m\ell$ and 0.006-0.024 $\mu\textrm{g}$/$m\ell$ both in the presence and absence of metabolic activation system, respectively. All 17 chemicals revealed no significant induction of chromosomal aberration both in the presence and absence of metabolic activation system in this assay. From the results of chromosomal aberration assay with 17 synthetic chemicals in Chinese hamster lung cells in vitro, we did not observed positive clastogenic results in this study.

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In Vitro Development and Apoptosis in Haploid, Diploid Parthenotes and Fertilized Embryos

  • Y. J Chung;Lee, H. Y.;S. H. Jun;X. S. Cui;Kim, N. H.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.28-28
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    • 2003
  • Haploid parthenotes have been shown to be developmentally delayed compared with diploid parthenogenetic embryos in the mouse and pig. These developmental defects have been hypothesized to rusult from insufficient parthenogenetic activation, suboptimal in vitro culture conditions, or genemic imprinting. In the present study we compared the incidence of apoptosis and apoptosis related gene expression in pig haploid, diploid parthenotes and fertilized embryos. In vitro matured porcine oocytes were activated by electrical stimulation. Haploid activated oocytes with two polar bodies under stereomicroscopy were defined haploid parthenotes, oocytes with one polar body were defined as diploid parthenotes after 3h cycloheximide teatment. The morphological analysis of apoptosis in embryos was carried out using propidium iodide staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. The expression of Bcl-xL, Bak and P53 in haploid, diploid and in vivo fertilized blastocysts was determined using RT-PCR. Lower number of the haploid pig parthenotes developed to the morulae and blastocysts compared to the diploid parthnotes. Number of cells significantly lower in the haploid-derived blastocysts than diploid-derived it. Developmentally retarded haploid parthenotes exibited apoptosis at a significantly higher frequency than did diploid parthenotes and fertilized embryos. Level of Bcl-xL expression, diploid parthenotes similar to in vivo-derived it was higher than haploid parthenotes. However, Bak and P53 mRNA expression were not different among haploid, diploid, and fertilized embryos. This result suggested that parthenogenetic activation and parthenogenesis themselves do not cause apoptosis, but haploid increases the incidence of apoptosis in preimplantation embryos. Apoptosis may be due to decrease expression of Bcl-xL in haploid parthenotes developing in vitro.

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Nuclear Maturation and Pronuclei Formation in Bovine Oocytes Matured In Vitro for Prolonged Period (체외 성숙 시간에 따른 소 난자의 처녀 발생)

  • 유형진;최승철;이상호
    • Korean Journal of Animal Reproduction
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    • v.17 no.4
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    • pp.331-337
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    • 1994
  • Response of the oocytes to parthenogenetic activation is one of the indice for cytoplasmic maturation. Maturational age-dependent parthenogenetic activation was examined in bovine oocytes. Follicular oocytes recovered from the slaughter house ovaries were matured in vitro in TCM 199+15% FCS+1Oiu/ml PMSG +10 iu/ml hCG from 24 to 48 h at 6 h intervals. The in vitro matured oocytes were activated by 7% ethanol for 7 min. The nuclear maturation and the cytoplasmic maturation were analysed by the nuclear configuration and pronuclei formation stained by rapid staining method. Cumulus oophori expansion increased as the maturation time increased. Proportions of the nuclear maturation were 81, 89, 72, 60 and 60% in IVM 24, 30, 36, 42 and 48 h groups, respectively. Abnor¬mality in metaphase II chromosome increased sharply from 36 h IVM. The rates of the pronuclei formation and diploid upon ethanol activation were 67, 68, 73, 84 and 87%, and 4, 5, 10, 16 and 20% in IVM 24, 30, 36, 42 and 48 h groups, respectively. It was suggested that maturational age increased the formation of the pronuclei and diploid, and that cytoplasmic maturation require longer maturation period than normal nuclear maturation. These results should be useful for determination of an appropriate time for fertilization in mammalian eggs matured or preincubated in vitro.

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Impact of In-vitro Fertility and Matrix Metalloproteinases Activation of Spermatozoa by Supplement of Tea-N-Tris to Sperm Cryopreservation of Miniature Pig (미니돼지 정자 동결 보존에 Tea-N-Tris의 첨가가 체외 수정 및 MMPs 활성에 미치는 영향)

  • Kim, Sang-Hwan;Kang, Hyun-Ah;Park, Yong-Su;Yoon, Jong-Taek
    • Journal of Embryo Transfer
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    • v.29 no.1
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    • pp.83-90
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    • 2014
  • The main purpose of this study is to estimate the effect of adding Tea-N-Tris (TES) to the freezing buffer for miniature pig sperm. In particular, we attempted to identify the association between the MMPs expression and the fertility and viability of frozen sperm from each extender (LEY (Lactose Egg-Yolk), TLE (TES + LEY), TFGE (TES + Fructose + Glucose Egg-Yolk)). In accordance with this, Hypoosmotic Swelling Test (HOST) respond test was the lowest among sperms frozen in LEY while the highest HOST respond was observed among sperms frozen in TLE. Furthermore, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of MMP-9 and MMP-2 was the highest in sperms frozen in LEY, Meanwhile, sperms from the TFGE and TLE group showed lower level of MMP-9 and MMP-2 expression in the order of TLE being the lowest. LEY group showed lower rate of blastocyst development than the TES supplement group, although the difference was not statistically significant. Meanwhile the rate of blastocyst development appeared similar when sperms from TLE and TFGE group were used for IVF. Together, these results indicate that adding Tea-N-Tris to the sperm freezing buffer only suppresses MMPs protein activation but also maximize in-vitro fertility, providing a means to improve the success rate in the in vitro manipulation of miniature pig sperm.